Three-dimensional perfusion culture of human bone marrow cells

¹ Departments of Surgery and of Research, Basel, Switzerland
² Department of Oncology, Biology and Genetics, Genova, Italy

^* To whom correspondence should be addressed. E-mail: imartin{at}uhbs.ch.

	Abstract

Three-dimensional (3D) culture systems are critical to investigate cell physiology and to engineer tissue grafts. Here we describe a simple yet innovative bioreactor-based approach to seed, expand and differentiate bone marrow stromal cells (BMSC) directly in a 3D environment, bypassing the conventional process of monolayer (2D) expansion. The system, based on the perfusion of bone marrow nucleated cells through porous 3D scaffolds, supported the formation of stromal-like tissues, where BMSC could be co-cultured with hematopoietic progenitor cells, in proportions dependent on the specific medium supplements. The resulting engineered constructs, when implanted ectopically in nude mice, generated bone tissue more reproducibly, uniformly and extensively than scaffolds loaded with 2D-expanded BMSC. The developed system may thus be used as a 3D in vitro model of bone marrow to study interactions between BMSC and hematopoietic cells, as well as to streamline manufacture of osteoinductive grafts in the context of regenerative medicine.